Patient-Derived Organoids for In Vivo Validation of In Vitro Data

Kdimati, Said; Bürtin, Florian; Linnebacher, Michael; Mullins, Christina Susanne

doi:10.1007/978-1-0716-2788-4_8

Cited by 2 publications

(3 citation statements)

References 52 publications

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“…Here, we leveraged gene editing in human keratinocytes and an organotypic epidermal model to elucidate the pathogenic mechanisms underlying a rare genetic disorder, which allowed us to identify much-needed potential therapeutic avenues. Though testing if our findings translate to an effective treatment approach for patients remains to be done, our work nevertheless underscores the power of organoids to generate large-scale transcriptomics and proteomics data sets that can be mined for rational drug targets to validate both in vitro and in patient-derived tissues (80,(82)(83)(84)(85).…”

Section: Discussionmentioning

confidence: 94%

Targeting SERCA2 in organotypic epidermis reveals MEK inhibition as a therapeutic strategy for Darier disease

Zaver¹,

Sarkar²,

Egolf³

et al. 2023

JCI Insight

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Mutation of the ATP2A2 gene encoding sarco-endoplasmic reticulum calcium ATPase 2 (SERCA2) was linked to Darier disease more than 2 decades ago; however, there remain no targeted therapies for this disorder causing recurrent skin blistering and infections. Since Atp2a2 -knockout mice do not phenocopy its pathology, we established a human tissue model of Darier disease to elucidate its pathogenesis and identify potential therapies. Leveraging CRISPR/Cas9, we generated human keratinocytes lacking SERCA2, which replicated features of Darier disease, including weakened intercellular adhesion and defective differentiation in organotypic epidermis. To identify pathogenic drivers downstream of SERCA2 depletion, we performed RNA sequencing and proteomics analysis. SERCA2-deficient keratinocytes lacked desmosomal and cytoskeletal proteins required for epidermal integrity and exhibited excess MAPK signaling, which modulates keratinocyte adhesion and differentiation. Immunostaining patient biopsies substantiated these findings, with lesions showing keratin deficiency, cadherin mislocalization, and ERK hyperphosphorylation. Dampening ERK activity with MEK inhibitors rescued adhesive protein expression and restored keratinocyte sheet integrity despite SERCA2 depletion or chemical inhibition. In sum, coupling multiomic analysis with human organotypic epidermis as a preclinical model, we found that SERCA2 haploinsufficiency disrupts critical adhesive components in keratinocytes via ERK signaling and identified MEK inhibition as a treatment strategy for Darier disease.

show abstract

Section: Discussionmentioning

confidence: 94%

Targeting SERCA2 in organotypic epidermis reveals MEK inhibition as a therapeutic strategy for Darier disease

Zaver¹,

Sarkar²,

Egolf³

et al. 2023

JCI Insight

View full text Add to dashboard Cite

show abstract

“…Here, we leveraged gene editing in human keratinocytes and an organotypic epidermal model to elucidate the pathogenic mechanisms underlying a rare genetic disorder, which allowed us to identify much-needed potential therapeutic avenues. Though testing if our findings translate to an effective treatment approach for patients remains to be done, our work nevertheless underscores the power of organoids to generate large-scale transcriptomic and proteomic datasets that can be mined for rational drug targets to validate both in vitro and in patient-derived tissues (77, 79–82).…”

Section: Discussionmentioning

confidence: 94%

“…The copyright holder for this preprint this version posted March 7, 2023. ; https://doi.org/10.1101/2023.03.07.531620 doi: bioRxiv preprint proteomic datasets that can be mined for rational drug targets to validate both in vitro and in patient-derived tissues (77,(79)(80)(81)(82).…”

Section: Discussionmentioning

confidence: 99%

Targeting SERCA2 in organotypic epidermis reveals MEK inhibition as a therapeutic strategy for Darier disease

Zaver

Sarkar

Egolf

et al. 2023

Preprint

View full text Add to dashboard Cite

Mutation of the ATP2A2 gene encoding sarco-endoplasmic reticulum calcium ATPase 2 (SERCA2) was linked to Darier disease more than two decades ago; however, there remain no targeted therapies for this disorder causing recurrent skin blistering and infections. Since Atp2a2 knockout mice do not phenocopy its pathology, we established a human tissue model of Darier disease to elucidate its pathogenesis and identify potential therapies. Leveraging CRISPR/Cas9, we generated human keratinocytes lacking SERCA2, which replicated features of Darier disease, including weakened intercellular adhesion and defective differentiation in organotypic epidermis. To identify pathogenic drivers downstream of SERCA2 depletion, we performed RNA sequencing and proteomic analysis. SERCA2-deficient keratinocytes lacked desmosomal and cytoskeletal proteins required for epidermal integrity and exhibited excess MAP kinase signaling, which modulates keratinocyte adhesion and differentiation. Immunostaining patient biopsies substantiated these findings with lesions showing keratin deficiency, cadherin mis-localization, and ERK hyper-phosphorylation. Dampening ERK activity with MEK inhibitors rescued adhesive protein expression and restored keratinocyte sheet integrity despite SERCA2 depletion or chemical inhibition. In sum, coupling multi-omic analysis with human organotypic epidermis as a pre-clinical model, we found that SERCA2 haploinsufficiency disrupts critical adhesive components in keratinocytes via ERK signaling and identified MEK inhibition as a treatment strategy for Darier disease.

show abstract

Patient-Derived Organoids for In Vivo Validation of In Vitro Data

Cited by 2 publications

References 52 publications

Targeting SERCA2 in organotypic epidermis reveals MEK inhibition as a therapeutic strategy for Darier disease

Targeting SERCA2 in organotypic epidermis reveals MEK inhibition as a therapeutic strategy for Darier disease

Targeting SERCA2 in organotypic epidermis reveals MEK inhibition as a therapeutic strategy for Darier disease

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